化痰祛湿活血方对脂多糖诱导的巨噬细胞焦亡形态的影响

刘鸣昊; 刘素彤; 尚东方; 张丽慧; 顾亚娇; 赵文霞

doi:10.3969/j.issn.1001-5256.2022.09.014

化痰祛湿活血方对脂多糖诱导的巨噬细胞焦亡形态的影响

DOI: 10.3969/j.issn.1001-5256.2022.09.014

河南中医药大学第一附属医院脾胃肝胆科，郑州 450000

基金项目:

国家自然科学基金项目 (81904154);

河南省科技攻关计划 (202102310168);

河南省重点研发与推广专项课题 (192102310425);

河南省中医药科学研究专项课题 (2019JDZX2051);

河南省科技攻关计划项目 (202102310495);

河南省特色骨干学科中医学学科建设项目 (STG-ZYXKY-2020024)

伦理学声明：本研究方案于2021年9月16日经由河南中医药大学伦理委员会审批，批号为YFYDW2021026，符合实验室动物管理与使用准则。

利益冲突声明：本研究不存在研究者、伦理委员会成员以及与公开研究成果有关的利益冲突。

作者贡献声明：刘鸣昊负责课题设计；刘素彤负责资料分析，撰写论文；尚东方、张丽慧、顾亚娇参与收集数据，修改论文；赵文霞负责拟定写作思路，指导撰写文章并最后定稿。

详细信息

通信作者:
赵文霞，zhao-wenxia@163.com

计量
- 文章访问数: 591
- HTML全文浏览量: 109
- PDF下载量: 32
- 被引次数: 0
出版历程
- 收稿日期: 2022-02-15
- 录用日期: 2022-03-17
- 出版日期: 2022-09-20

Effect of Huatan Qushi Huoxue prescription on lipopolysaccharide-induced pyroptosis of macrophages

Department of Hepatology and Spleen-Stomach, The First Affiliated Hospital of Henan University of Traditional Chinese Medicine, Zhengzhou 450000, China

Research funding:

National Natural Science Foundation of China (81904154);

Science and Technology Research Program of Henan Province (202102310168);

Key RESEARCH and Promotion Project of Henan Province (192102310425);

Traditional Chinese Medicine Science Research Project of Henan Province (2019JDZX2051);

Key Science and Technology Project of Henan Province (202102310495);

TCM Discipline Construction Project of Characteristic Backbone Disciplines of Henan Province (STG-ZYXKY-2020024)

More Information

Corresponding author: ZHAO Wenxia, zhao-wenxia@163.com(ORCID: 0000-0001-6666-9469)

摘要

摘要: 目的化痰祛湿活血方对脂多糖(LPS)诱导的RAW264.7细胞焦亡形态的影响及机制研究。方法复制体外LPS诱导活化RAW264.7细胞模型，分为空白组，模型组，化痰祛湿活血方高、中、低剂量组，对照组。经相应的含药血清干预24 h。扫描电子显微镜观察各组细胞形态。免疫荧光对GSDMD-N进行定量定位分析。结果空白组细胞形态圆形规整，表面光滑；对照组细胞肿胀，表面有褶皱，包膜有隙，符合细胞焦亡形态。对照组细胞表面有气泡状突出物，伪足明显。细胞膜有孔隙，高剂量组细胞形态无肿胀，表面粗糙有伪足，胞膜上未见明显孔隙。低剂量和中剂量组细胞肿胀，细胞膜表面粗糙有孔隙，有伪足伸出。免疫荧光结果显示，与空白组相比模型组GSDMD-N阳性染色强度明显增加，对照组与中药组较模型组GSDMD-N阳性染色强度均有减弱。结论化痰祛湿活血方可改善巨噬细胞焦亡现象，减少GSDMD-N表达。
- 脂肪肝 /
- 巨噬细胞 /
- 细胞焦亡
Abstract: Objective To investigate the effect of Huatan Qushi Huoxue prescription on lipopolysaccharide (LPS)-induced pyroptosis of RAW264.7 cells and its mechanism. Methods An in vitro cell model of LPS-induced activated RAW264.7 was established and divided into blank group, model group, high-, middle-, and low-dose Huatan Qushi Huoxue prescription groups, and control group. The corresponding drug-containing serum intervention was performed for 24 hours. A scanning electron microscope was used to observe cell morphology, and immunofluorescence assay was used to perform quantitative localization of GSDMD-N. Results The cells in the blank group were round and regular in shape with smooth surface, and those in the control group were swollen, with folds on the surface and gaps in the capsule, which were consistent with the morphology of cell pyroptosis. The cells in the control group had bubbles on the surface with obvious pseudopodia and pores in cell membrane, and those in the high-dose group were not swollen and had a rough surface with pseudopodia, with no obvious pores in cell membrane. The cells in the low- and middle-dose groups were swollen and had a rough surface of cell membrane with pores and pseudopodia. Immunofluorescence assay showed that compared with the blank group, the model group had a significant increase in the positive staining intensity of GSDMD-N, and compared with the model group, the control group and the Traditional Chinese medicine group had a reduction in the positive staining intensity of GSDMD-N. Conclusion Huatan Qushi Huoxue prescription can improve the pyroptosis of macrophages and reduce the expression of GSDMD-N.
- Fatty Liver /
- Macrophages /
- Pyroptosis

HTML全文

图 1 电镜下巨噬细胞形态变化(低倍，×1500；高倍，×6000)

Figure 1. Morphological changes of macrophages under electron microscopy (low fold, ×1500; High fold, ×6000)

下载: 全尺寸图片幻灯片

图 2 巨噬细胞内GSDMD-N水平定位(×400)

注：蓝色代表PI染色结果；绿色代表GSDMD-N染色结果。

Figure 2. GSDMD-N level localization in macrophages(×400)

下载: 全尺寸图片幻灯片

参考文献(7)

[1]	YOUNOSSI Z, STEPANOVA M, ONG JP, et al. Nonalcoholic steatohepatitis is the fastest growing cause of hepatocellular carcinoma in liver transplant candidates[J]. Clin Gastroenterol Hepatol, 2019, 17(4): 748-755. e3. DOI: 10.1016/j.cgh.2018.05.057.
[2]	LANTHIER N. Targeting Kupffer cells in non-alcoholic fatty liver disease/non-alcoholic steatohepatitis: Why and how?[J]. World J Hepatol, 2015, 7(19): 2184-2188. DOI: 10.4254/wjh.v7.i19.2184.
[3]	SHANG DF, LIU MH, ZHANG LH, et al. Study on the effect of cell pyrodeath on nonalcoholic steatohepatitis based on the theory of "Treating liver from lung"[J]. Chin J Basic Med Tradit Chin Med, 2022, 28(4): 582-585. DOI: 10.19945/j.cnki.issn.1006-3250.20211014.001. 尚东方, 刘鸣昊, 张丽慧, 等. 基于"从肺治肝"理论探讨干预细胞焦亡对非酒精性脂肪性肝炎的影响[J]. 中国中医基础医学杂志, 2022, 28(4): 582-585. DOI: 10.19945/j.cnki.issn.1006-3250.20211014.001.
[4]	ZHANG LH, ZHANG JB, ZHAO WX. Effect of Huatan Qushi Huoxue prescription on the key proteins of the ADPN/PI3K/Akt signaling pathway in ratswith nonalcoholic steatohepatitis[J]. J Clin Hepatol, 2020, 36(4): 835-839. DOI: 10.3969/j.issn.1001-5256.2020.04.025. 张丽慧, 张剑波, 赵文霞. 化痰祛湿活血方对非酒精性脂肪性肝炎大鼠模型ADPN/PI3K/AKT通路关键蛋白的影响[J]. 临床肝胆病杂志, 2020, 36(4): 835-839. DOI: 10.3969/j.issn.1001-5256.2020.04.025.
[5]	REID DT, MCDONALD B, KHALID T, et al. Unique microbial-derived volatile organic compounds in portal venous circulation in murine non-alcoholic fatty liver disease[J]. Biochim Biophys Acta, 2016, 1862(7): 1337-1344. DOI: 10.1016/j.bbadis.2016.04.005.
[6]	BOARU SG, BORKHAM-KAMPHORST E, TIHAA L, et al. Expression analysis of inflammasomes in experimental models of inflammatory and fibrotic liver disease[J]. J Inflamm (Lond), 2012, 9(1): 49. DOI: 10.1186/1476-9255-9-49.
[7]	XU B, JIANG M, CHU Y, et al. Gasdermin D plays a key role as a pyroptosis executor of non-alcoholic steatohepatitis in humans and mice[J]. J Hepatol, 2018, 68(4): 773-782. DOI: 10.1016/j.jhep.2017.11.040.